1. Field of the Invention
This invention relates to monoclonal antibodies produced by hybridoma cell lines created by the fusion of myeloma cell lines with lymphocytes.
The invention further relates to methods for detecting anaerobic microorganisms in clinical samples; more specifically the present invention relates to methods for detecting Bacteroides species, common etiological agents in human anaerobic infections (Gorbach and Bartlett, New England Journal of Medicine, 290, 1177, 1237, 1289 (1974)).
It has now been found that the fusion of a mouse myeloma cell line, X63-Ag8.653, with a BALB/c mouse lymphocyte creates a hybridoma which produces a monoclonal antibody which is capable of specifically detecting 14 strains of Bacteroides gingivalis.
2. Discussion of the Prior Art
The black-pigmented oral anaerobe Bacteroides gingivalis has been increasingly implicated as an etiologic agent of severe periodontitis in adults (Slots, Journal of Dental Research, 63, 412 (1984)). Therefore, the detection of this anaerobic micro-organism in a clinical sample may serve as an important diagnostic indicator. Because of the difficulty and amount of time needed to culture, isolate and identify B. gingivalis from clinical specimens, serological methods would greatly aid in its detection. Before the use of hybridoma to produce monoclonal antibodies, species specific antibodies were made by adsorbing polyclonal animal sera with cross-reacting antigens. The use of an enzyme linked immunosorbent assay as a rapid serological method of identifying oral species of Bacteroides is described by Ebersole, et al., Journal of Clinical Microbiology, 19, 639 (1984). However, antisera prepared in animals are heterogeneous, unpredictable, and very limited in supply. The advent of a classical technique to produce monoclonal antibodies (Kohler and Milstein, Nature, 256, 495 (1975)) has allowed for the generation of immunodiagnostic reagents which are highly specific, homogeneous, and unlimited in supply.
A simplified technique based on the Kohler and Milstein methodology for creating hybridomas that produce monoclonal antibodies was described by Fazekas de St. Groth and Scheidegger (Journal of Immunological Methods, 35, 1 (1980)). The conditions described by Fazekas de St. Groth and Scheidegger have been utilized frequently; these conditions were modified to obtain the hybridoma producing the monoclonal antibody of the present invention (Simonson, et al., Journal of Dental Research, 65, 95 (1986)).
Recently, Strosberg, et al. disclosed in U.S. Pat. No. 4,780,407 a process for the immunological determination of Legionella pneumophilia bacteria using a monoclonal antibody produced by a hybridoma cell-line.